Generation of Human Fab Libraries for Phage Display
This protocol describes the generation of human antibody libraries in Fab format from 2.5 × 107 human peripheral blood or bone marrow mononuclear cells for their subsequent selection by phage display. Although it can be applied to the mining of both human naïve and immune antibody repertoires, the procedure is primarily intended for the generation of fully human monoclonal antibodies from patients with endogenous antibody responses of interest and limited availability of clinical specimens.
Key wordsImmune antibody repertoires Antibody fragment Antibody libraries Fab Human monoclonal antibodies Library Phage display Phagemid
This work was supported by the Intramural Research Program of the Center for Cancer Research, National Cancer Institute, National Institutes of Health. I thank current and former members of my laboratory, in particular Dr. Ka Yin Kwong, Dr. Jiahui Yang, Dr. Sivasubramanian Baskar, and Jessica M. Suschak, for their contributions to this protocol.
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